Citation: YANG Lin, LI Guo-Qing, HUANG Bao-Yang, WANG Quan-Zhong, LONG Qi-Xin, WANG Xun-Zhang. Solubilization Analysis of GST Fusion Protein Expressed in Baculovirus System .VIROLOGICA SINICA, 2000, 15(2) : 143-148.

Solubilization Analysis of GST Fusion Protein Expressed in Baculovirus System

  • Available online: 05 June 2000
  • Insect Sf 9 cells were infected with constructed recombinant virus AcMNPV OCC - GST 6xHis Etp 28, which could produce GST fusion protein and the supernatant of 72 h pi cell lysate was examined by SDS PAGE. The results showed that GST 6xHis Etp 28 fusion protein of 53 kDa was expressed in insoluble status. After the sodium salt of the alkylanionic detergent sarkosyl was added to insect cell lysis buffer to a final concentration of 1.5%, and the final concentration of TritonX 100 was increased from 1% to 2%, at least 1/3 of GST 6xHis Etp 28 appeared to be soluble, which was examined by SDS PAGE. Then soluble GST 6xHis Etp 28 was purified by affinity chromatography using glutathione agarose.

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    Solubilization Analysis of GST Fusion Protein Expressed in Baculovirus System

    • 1. State key Lab for Biological Control,Biopharmaceutical Center,Zhangshan University,Guangzhou 510275,China

    Abstract: Insect Sf 9 cells were infected with constructed recombinant virus AcMNPV OCC - GST 6xHis Etp 28, which could produce GST fusion protein and the supernatant of 72 h pi cell lysate was examined by SDS PAGE. The results showed that GST 6xHis Etp 28 fusion protein of 53 kDa was expressed in insoluble status. After the sodium salt of the alkylanionic detergent sarkosyl was added to insect cell lysis buffer to a final concentration of 1.5%, and the final concentration of TritonX 100 was increased from 1% to 2%, at least 1/3 of GST 6xHis Etp 28 appeared to be soluble, which was examined by SDS PAGE. Then soluble GST 6xHis Etp 28 was purified by affinity chromatography using glutathione agarose.

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